A Full Exploitation of Nature's Medicine - Apple

Plants are the original and the most effective medicines.

Used since antiquity for the prevention and treatment of numerous diseases, medicinal plants
represent the natural, miraculous and special medicine that eases pain, heals illnesses and
reestablish the harmony between human beings and Nature.
The apple has a long history. We can read about it in the Bible and in many other religious
books, such as the Egyptian Book of the Dead. Its symbolism is present in numerous tales and
legends. The apple played an important part in certain rites.
Apple extracts fight cancer, due to proantocians, powerful natural antioxidants [1]. Because of
the polyphenols it contains, especially quercetin - the most powerful antioxidant derived from
plants - it helps eliminate sebum and consolidate skin immunity, it eliminates roughness,
smooths tired skin and has an antiaging effect [2].
The aim of this paper is to contribute to obtaining better apple extracts from dried and minced
fruit, by reusing the pre-extracted vegetal material, by combining acetone and ethanol
extraction and minimizing the quantity of resources used. The raw material is the Malus
sylvestris fructus apple, minced and dried.

We have chosen these extraction methods in order to fully characterize raw materials,
considering that extracted fractions can be used, in the future, in cosmetic creams with
antiaging effect.
The process takes place, three fractions containing hydroxiacids, saccharides,
unsaturated fats (fr.1), polyphenols, proantocians, flavonoids (fr.2) and pectines (fr.3) are
collected and characterised. These fractions are characterised qualitatively and quantitatively,
via spectrophotometric analysis, thin layer chromatography and gas chromatography coupled
with mass spectrometry [3]. The results from the sample were obtained by comparing the
mass spectrum obtained with the one in the Nist spectral library.

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Identification and quantification techniques used:

The caffeic acid content was determined via gas
chromatography coupled with mass spectrometry after
being
derivatized
with
bis(trimethylsilyl)trifluoroacetamide (BSTFA).
The UV chromatogram for indirect marking of
proantociani compounds found in the Malus sylvestris
extract.
Absorbance is read with a spectrophotometer at 660 nm in the 1 cm tray and compared with
the 20% solution of sodium carbonate (R), taken as reference; the device used was the
Shimadzu 1205 UV-VIS spectrophotometer.

Obtaining the F1 fraction - The 1.7 liters of extract obtained were subject to vacuum
concentration (556-500 mbar) and an evaporation temperature of 30-400C, while the active
components were protected until the dry residue was obtained. The F1 fraction obtained,
amounting to 10.2 g, is a red-brown paste, subject to gas chromatography coupled with mass
spectrometry. The analysis underlined the presence of large quantities of: hydroxiacids, of
which the malic acid is the most important 45.32%, a crucial component in the Krebs cycle
for ATP formation; 2-keto-gluconic acid 11.24%- involved in the synthesis of vitamin C;
numerous saccharides: lyxosazone, fructose, altrose, glucopyranose, galactopyranose,
methyl-glucofuranoside, glucitol, glucopyranosis varying between 0.1 and 5.3% ;
polyunsaturated fatty acids: linoleic 0,518% and oleic -9 monounsaturated 0.553%;

Obtaining the F2 fraction - The vegetal product - 88 g resulting from the previous stage
(Soxhlet extraction) was pressed and subjected to successive ethanol extraction 80%, 70%,
60% and 50%, in order to isolate the phenolic compounds: polyphenol carboxylic acids,
flavonoids and proantocians.
The 3.4 liters of extract obtained was subject to vacuum concentration (175-200 mbar) and to
steam at a temperature of 30-40 0C, then conditioned via inclusion in propylene glycol at a 1:1
ratio of plant : final extract. The 100 ml of F2 fraction conditioned in propylenglycol is a
reddish, clear liquid subject to thin layer chromatographic analysis in order to identify
phenolic components: polyphenol carboxylic acids, proantocians, spectrophotometric
determination of the quantity of polyphenol compounds expressed in caffeic acid, of
flavonoids expressed in rutin and of proantocians and of caffeic acid via GC-MS.
The concentration of caffeic acid obtained in the extractive solution II is of 0.13 mg/100ml of
Malus sylvestris extract.

Obtaining the F3 fraction - The vegetal product - 70 g, the previous step was pressed and
subjected to further extraction with hot distilled water - 60 C in a laboratory homogenizer
sheathed heating and variable speed in three steps. Extraction time was 30 minutes for each
step. Report plant product: extraction solvent was 1: 30. Extract obtained in an amount of 1.8
L was subjected to concentration under vacuum (75-100 mbar) and vapor temperature 30[Tastai text]

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400C up to a ratio 1:20; product obtained as a viscous syrup was poured into thin jet in
pharmaceutical ethanol by stirring, at which the precipitation occurred soluble fraction F3,
which are separated by Buchner filtration and dried. Achieved a pectin type polysaccharide
prepared in quantity. 0.6 g/100g plant product, with the ability to form gels. F3 fraction was
analyzed by GC-MS after acid hydrolysis, to highlight oze components. Identification of
compounds was achieved by comparing mass spectra with library spectra from NIST. From a
total of 25 separate 22 compounds were identified. Note that malic acid and keto-gluconic
acid is present at a level of 50% of the separated compounds namely 45.32% respectively
15.37%. Significant content in the mixture of compounds separated is presents of 4%
Pregnan. Also, glucose (in different isoforms) is represented in a relatively high content of
approx. 12%.

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